Enhanced Synaptic-Related Transcriptional Programs in Human Forebrain Organoids for Modeling Synaptopathy-Associated Neurodevelopmental Disorders
- 1Department of Pharmacology, Second Military Medical University (Naval Medical University), Shanghai 200433, China
Abstract
Human cerebral organoids (COs) are well-recognized in vitro models for recapitulating human fetal brain development. However, spontaneous unguided differentiation of COs leads to severe regional heterogeneity, limiting standardized research and high-throughput drug screening. In this study, we generated optimized human forebrain organoids (FBs) using a triple SMAD/Wnt inhibition strategy modified from classical CO culture systems. Immunofluorescence staining and bulk RNA sequencing validated the physiological authenticity of FBs and elucidated prominent phenotypic differences between FBs and COs. Molecular analyses confirmed robust and specific forebrain specification in FBs, characterized by sustained expression of canonical forebrain markers and dominant enrichment of neocortical and subpallial signatures, while posterior brain lineage programs were markedly suppressed. Furthermore, FBs faithfully recapitulated diverse fetal forebrain cellular types and rudimentary cortical laminar structures. Transcriptomic profiling revealed that FBs exhibited a marked transcriptional shift toward gene sets associated with neuronal differentiation and synaptic-related biological processes. Gene sets linked to synaptic assembly, transmission, and plasticity were transcriptionally up-regulated, whereas cell-cycle- and proliferation-associated transcriptional programs were downregulated as compared to COs. In conclusion, the optimized FBs faithfully recapitulated key human forebrain developmental features and exhibited enhanced synapse-related transcriptional signatures relative to COs. This refined organoid system provides a valuable in vitro platform for studying forebrain circuit ontogeny and dissecting the molecular pathogenesis of synaptopathy-related neurodevelopmental disorders.
Related
- Isogenic forebrain organoids uncover early neurodevelopmental alterations and imbalances in neuronal function leading to hyperexcitation in Gaucher disease
- Concussive Injury in Human Forebrain Organoids Alters Neurogenesis and Neuroimmune Signaling
- Krüppel-like factors KLF5 & KLF8 emerge as potential master transcriptional regulators of Alzheimer's disease, as revealed on cell fate regulomes in human brain organoids
- Immunometabolic synaptopathy: linking metabolic syndrome to epileptogenesis through trained immunity and parvalbumin interneuron vulnerability
- Stage-specific proteomic signatures of synaptopathy in Alzheimers disease
BibTeX
@article{zhang2026enhanced,
title = {{Enhanced Synaptic-Related Transcriptional Programs in Human Forebrain Organoids for Modeling Synaptopathy-Associated Neurodevelopmental Disorders}},
author = {Xiu-Ping Zhang and Ling-Liang Zhang and Xi-Yuan Wang and Shu-Na Wang and Chao-Yu Miao},
journal = {Cells},
year = {2026},
month = oct,
doi = {10.3390/cells15201842},
url = {https://www.mdpi.com/2073-4409/15/20/1842},
note = {CC BY 4.0},
}
RIS
TY - JOUR
TI - Enhanced Synaptic-Related Transcriptional Programs in Human Forebrain Organoids for Modeling Synaptopathy-Associated Neurodevelopmental Disorders
AU - Xiu-Ping Zhang
AU - Ling-Liang Zhang
AU - Xi-Yuan Wang
AU - Shu-Na Wang
AU - Chao-Yu Miao
PY - 2026
DA - 2026/10/11
JO - Cells
DO - 10.3390/cells15201842
UR - https://www.mdpi.com/2073-4409/15/20/1842
AB - Human cerebral organoids (COs) are well-recognized in vitro models for recapitulating human fetal brain development. However, spontaneous unguided differentiation of COs leads to severe regional heterogeneity, limiting standardized research and high-throughput drug screening. In this study, we generated optimized human forebrain organoids (FBs) using a triple SMAD/Wnt inhibition strategy modified from classical CO culture systems. Immunofluorescence staining and bulk RNA sequencing validated the physiological authenticity of FBs and elucidated prominent phenotypic differences between FBs and COs. Molecular analyses confirmed robust and specific forebrain specification in FBs, characterized by sustained expression of canonical forebrain markers and dominant enrichment of neocortical and subpallial signatures, while posterior brain lineage programs were markedly suppressed. Furthermore, FBs faithfully recapitulated diverse fetal forebrain cellular types and rudimentary cortical laminar structures. Transcriptomic profiling revealed that FBs exhibited a marked transcriptional shift toward gene sets associated with neuronal differentiation and synaptic-related biological processes. Gene sets linked to synaptic assembly, transmission, and plasticity were transcriptionally up-regulated, whereas cell-cycle- and proliferation-associated transcriptional programs were downregulated as compared to COs. In conclusion, the optimized FBs faithfully recapitulated key human forebrain developmental features and exhibited enhanced synapse-related transcriptional signatures relative to COs. This refined organoid system provides a valuable in vitro platform for studying forebrain circuit ontogeny and dissecting the molecular pathogenesis of synaptopathy-related neurodevelopmental disorders.
SN - 2073-4409
ER -